Rapid and simultaneous quantification of viable Escherichia coli O157:H7 and Salmonella spp. in milk through multiplex real-time PCR
Escherichia coli O157:H7 and Salmonella spp. in milk are 2 common pathogens that cause foodborne diseases. An accurate, rapid, specific method has been developed for the simultaneous detection of viable E. coli O157:H7 and Salmonella spp. in milk. Two specific genes, namely, fliC from E. coli O157:H...
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| Veröffentlicht in: | Journal of dairy science 2017-11, Vol.100 (11), p.8804-8813 |
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| creator | Zhou, Baoqing Liang, Taobo Zhan, Zhongxu Liu, Rui Li, Fan Xu, Hengyi |
| description | Escherichia coli O157:H7 and Salmonella spp. in milk are 2 common pathogens that cause foodborne diseases. An accurate, rapid, specific method has been developed for the simultaneous detection of viable E. coli O157:H7 and Salmonella spp. in milk. Two specific genes, namely, fliC from E. coli O157:H7 and invA from Salmonella spp., were selected to design primers and probes. A combined treatment containing sodium deoxycholate (SDO) and propidium monoazide (PMA) was applied to detect viable E. coli O157:H7 and Salmonella spp. only. Traditional culture methods and SDO-PMA-multiplex real-time (mRT) PCR assay were applied to determine the number of viable E. coli O157:H7 and Salmonella spp. in cell suspensions with different proportions of dead cells. These methods revealed consistent findings regarding the detected viable cells. The detection limit of the SDO-PMA-mRT-PCR assay reached 102 cfu/mL for Salmonella spp. and 102 cfu/mL for E. coli O157:H7 in milk. The detection limit of SDO-PMA-mRT-PCR for E. coli O157:H7 and Salmonella spp. in milk was significantly similar even in the presence of 106 cfu/mL of 2 nontarget bacteria. The proposed SDO-PMA-mRT-PCR assay is a potential approach for the accurate and sensitive detection of viable E. coli O157:H7 and Salmonella spp. in milk. |
| doi_str_mv | 10.3168/jds.2017-13362 |
| format | Article |
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An accurate, rapid, specific method has been developed for the simultaneous detection of viable E. coli O157:H7 and Salmonella spp. in milk. Two specific genes, namely, fliC from E. coli O157:H7 and invA from Salmonella spp., were selected to design primers and probes. A combined treatment containing sodium deoxycholate (SDO) and propidium monoazide (PMA) was applied to detect viable E. coli O157:H7 and Salmonella spp. only. Traditional culture methods and SDO-PMA-multiplex real-time (mRT) PCR assay were applied to determine the number of viable E. coli O157:H7 and Salmonella spp. in cell suspensions with different proportions of dead cells. These methods revealed consistent findings regarding the detected viable cells. The detection limit of the SDO-PMA-mRT-PCR assay reached 102 cfu/mL for Salmonella spp. and 102 cfu/mL for E. coli O157:H7 in milk. The detection limit of SDO-PMA-mRT-PCR for E. coli O157:H7 and Salmonella spp. in milk was significantly similar even in the presence of 106 cfu/mL of 2 nontarget bacteria. The proposed SDO-PMA-mRT-PCR assay is a potential approach for the accurate and sensitive detection of viable E. coli O157:H7 and Salmonella spp. in milk.</description><identifier>ISSN: 0022-0302</identifier><identifier>EISSN: 1525-3198</identifier><identifier>DOI: 10.3168/jds.2017-13362</identifier><identifier>PMID: 28865862</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Azides ; Cattle ; DNA Primers ; Escherichia coli O157 - isolation & purification ; Food Microbiology ; foodborne pathogen ; Milk - microbiology ; Multiplex Polymerase Chain Reaction - methods ; Multiplex Polymerase Chain Reaction - veterinary ; multiplex real-time PCR ; Propidium - analogs & derivatives ; propidium monoazide ; Real-Time Polymerase Chain Reaction - methods ; Salmonella - genetics ; Salmonella - isolation & purification ; Sensitivity and Specificity ; sodium deoxycholate</subject><ispartof>Journal of dairy science, 2017-11, Vol.100 (11), p.8804-8813</ispartof><rights>2017 American Dairy Science Association</rights><rights>Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c2992-86a45f9c89c4e9b5c04f1c6ebb0fa555e6129d4cc9778fa02069c43b7bfd1bcf3</citedby><cites>FETCH-LOGICAL-c2992-86a45f9c89c4e9b5c04f1c6ebb0fa555e6129d4cc9778fa02069c43b7bfd1bcf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.3168/jds.2017-13362$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>315,781,785,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28865862$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhou, Baoqing</creatorcontrib><creatorcontrib>Liang, Taobo</creatorcontrib><creatorcontrib>Zhan, Zhongxu</creatorcontrib><creatorcontrib>Liu, Rui</creatorcontrib><creatorcontrib>Li, Fan</creatorcontrib><creatorcontrib>Xu, Hengyi</creatorcontrib><title>Rapid and simultaneous quantification of viable Escherichia coli O157:H7 and Salmonella spp. in milk through multiplex real-time PCR</title><title>Journal of dairy science</title><addtitle>J Dairy Sci</addtitle><description>Escherichia coli O157:H7 and Salmonella spp. in milk are 2 common pathogens that cause foodborne diseases. An accurate, rapid, specific method has been developed for the simultaneous detection of viable E. coli O157:H7 and Salmonella spp. in milk. Two specific genes, namely, fliC from E. coli O157:H7 and invA from Salmonella spp., were selected to design primers and probes. A combined treatment containing sodium deoxycholate (SDO) and propidium monoazide (PMA) was applied to detect viable E. coli O157:H7 and Salmonella spp. only. Traditional culture methods and SDO-PMA-multiplex real-time (mRT) PCR assay were applied to determine the number of viable E. coli O157:H7 and Salmonella spp. in cell suspensions with different proportions of dead cells. These methods revealed consistent findings regarding the detected viable cells. The detection limit of the SDO-PMA-mRT-PCR assay reached 102 cfu/mL for Salmonella spp. and 102 cfu/mL for E. coli O157:H7 in milk. The detection limit of SDO-PMA-mRT-PCR for E. coli O157:H7 and Salmonella spp. in milk was significantly similar even in the presence of 106 cfu/mL of 2 nontarget bacteria. The proposed SDO-PMA-mRT-PCR assay is a potential approach for the accurate and sensitive detection of viable E. coli O157:H7 and Salmonella spp. in milk.</description><subject>Animals</subject><subject>Azides</subject><subject>Cattle</subject><subject>DNA Primers</subject><subject>Escherichia coli O157 - isolation & purification</subject><subject>Food Microbiology</subject><subject>foodborne pathogen</subject><subject>Milk - microbiology</subject><subject>Multiplex Polymerase Chain Reaction - methods</subject><subject>Multiplex Polymerase Chain Reaction - veterinary</subject><subject>multiplex real-time PCR</subject><subject>Propidium - analogs & derivatives</subject><subject>propidium monoazide</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Salmonella - genetics</subject><subject>Salmonella - isolation & purification</subject><subject>Sensitivity and Specificity</subject><subject>sodium deoxycholate</subject><issn>0022-0302</issn><issn>1525-3198</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kE1v1DAQhi0EokvhyhH5yCWLP-LE5oZWhSJVKipwthxnzE5x4tROqnLnh5PdLdw4jUZ65p2Zh5DXnG0lb_S7275sBeNtxaVsxBOy4UqoSnKjn5INY0JUTDJxRl6Ucru2XDD1nJwJrRulG7Ehv2_chD11Y08LDkuc3QhpKfRuceOMAb2bMY00BXqProtAL4rfQ0a_R0d9ikivuWrfX7bHiK8uDmmEGB0t07SlONIB408673NafuzpYQFOER5oBherGQegX3Y3L8mz4GKBV4_1nHz_ePFtd1ldXX_6vPtwVXlhjKh042oVjNfG12A65VkduG-g61hwSilouDB97b1pWx0cE6xZSdm1Xeh554M8J29PuVNOdwuU2Q5Y_OHc49OWG6mkYbXQK7o9oT6nUjIEO2UcXP5lObMH83Y1bw_m7dH8OvDmMXvpBuj_4X9Vr4A-AbB-eI-QbfEIo4ceM_jZ9gn_l_0Hip6TCQ</recordid><startdate>201711</startdate><enddate>201711</enddate><creator>Zhou, Baoqing</creator><creator>Liang, Taobo</creator><creator>Zhan, Zhongxu</creator><creator>Liu, Rui</creator><creator>Li, Fan</creator><creator>Xu, Hengyi</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>201711</creationdate><title>Rapid and simultaneous quantification of viable Escherichia coli O157:H7 and Salmonella spp. in milk through multiplex real-time PCR</title><author>Zhou, Baoqing ; Liang, Taobo ; Zhan, Zhongxu ; Liu, Rui ; Li, Fan ; Xu, Hengyi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2992-86a45f9c89c4e9b5c04f1c6ebb0fa555e6129d4cc9778fa02069c43b7bfd1bcf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Animals</topic><topic>Azides</topic><topic>Cattle</topic><topic>DNA Primers</topic><topic>Escherichia coli O157 - isolation & purification</topic><topic>Food Microbiology</topic><topic>foodborne pathogen</topic><topic>Milk - microbiology</topic><topic>Multiplex Polymerase Chain Reaction - methods</topic><topic>Multiplex Polymerase Chain Reaction - veterinary</topic><topic>multiplex real-time PCR</topic><topic>Propidium - analogs & derivatives</topic><topic>propidium monoazide</topic><topic>Real-Time Polymerase Chain Reaction - methods</topic><topic>Salmonella - genetics</topic><topic>Salmonella - isolation & purification</topic><topic>Sensitivity and Specificity</topic><topic>sodium deoxycholate</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhou, Baoqing</creatorcontrib><creatorcontrib>Liang, Taobo</creatorcontrib><creatorcontrib>Zhan, Zhongxu</creatorcontrib><creatorcontrib>Liu, Rui</creatorcontrib><creatorcontrib>Li, Fan</creatorcontrib><creatorcontrib>Xu, Hengyi</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of dairy science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhou, Baoqing</au><au>Liang, Taobo</au><au>Zhan, Zhongxu</au><au>Liu, Rui</au><au>Li, Fan</au><au>Xu, Hengyi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rapid and simultaneous quantification of viable Escherichia coli O157:H7 and Salmonella spp. in milk through multiplex real-time PCR</atitle><jtitle>Journal of dairy science</jtitle><addtitle>J Dairy Sci</addtitle><date>2017-11</date><risdate>2017</risdate><volume>100</volume><issue>11</issue><spage>8804</spage><epage>8813</epage><pages>8804-8813</pages><issn>0022-0302</issn><eissn>1525-3198</eissn><abstract>Escherichia coli O157:H7 and Salmonella spp. in milk are 2 common pathogens that cause foodborne diseases. An accurate, rapid, specific method has been developed for the simultaneous detection of viable E. coli O157:H7 and Salmonella spp. in milk. Two specific genes, namely, fliC from E. coli O157:H7 and invA from Salmonella spp., were selected to design primers and probes. A combined treatment containing sodium deoxycholate (SDO) and propidium monoazide (PMA) was applied to detect viable E. coli O157:H7 and Salmonella spp. only. Traditional culture methods and SDO-PMA-multiplex real-time (mRT) PCR assay were applied to determine the number of viable E. coli O157:H7 and Salmonella spp. in cell suspensions with different proportions of dead cells. These methods revealed consistent findings regarding the detected viable cells. The detection limit of the SDO-PMA-mRT-PCR assay reached 102 cfu/mL for Salmonella spp. and 102 cfu/mL for E. coli O157:H7 in milk. The detection limit of SDO-PMA-mRT-PCR for E. coli O157:H7 and Salmonella spp. in milk was significantly similar even in the presence of 106 cfu/mL of 2 nontarget bacteria. The proposed SDO-PMA-mRT-PCR assay is a potential approach for the accurate and sensitive detection of viable E. coli O157:H7 and Salmonella spp. in milk.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>28865862</pmid><doi>10.3168/jds.2017-13362</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Azides Cattle DNA Primers Escherichia coli O157 - isolation & purification Food Microbiology foodborne pathogen Milk - microbiology Multiplex Polymerase Chain Reaction - methods Multiplex Polymerase Chain Reaction - veterinary multiplex real-time PCR Propidium - analogs & derivatives propidium monoazide Real-Time Polymerase Chain Reaction - methods Salmonella - genetics Salmonella - isolation & purification Sensitivity and Specificity sodium deoxycholate |
| title | Rapid and simultaneous quantification of viable Escherichia coli O157:H7 and Salmonella spp. in milk through multiplex real-time PCR |
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